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机构地区:[1]暨南大学附属第一医院心内科,广州510630 [2]暨南大学医学院组织学与胚胎学系
出 处:《临床心血管病杂志》2013年第6期470-474,共5页Journal of Clinical Cardiology
基 金:广东省科技计划项目(No:2010-1096-136);广州市科技计划项目(No:2010Y1-C941);暨南大学第一临床医学院重点学科基金项目(No:2010-4)
摘 要:目的:研究不同浓度辛伐他汀对体外培养的人脐静脉血晚期内皮祖细胞(EPCs)数量和功能的影响。方法:用密度梯度离心法分离人脐静脉血单个核细胞,接种于人纤维连接蛋白包被的培养瓶,在EGM-2MV培养基中诱导分化获得晚期EPCs,并通过流式细胞术及免疫荧光染色进行鉴定。取生长状态良好的第2代晚期EPCs与不同浓度辛伐他汀(10-8、10-7、10-6及10-5mol/L)培养24、48及72h后,采用CCK-8增殖能力检测、粘附能力测定及血管形成实验来观察辛伐他汀对晚期EPCs增殖、粘附及血管形成能力的影响。结果:从人脐静脉血中成功分离并培养出晚期EPCs。与对照组相比,较低浓度辛伐他汀(10-8、10-7与10-6mol/L)对晚期EPCs的增殖、粘附及血管形成有明显促进作用,以10-7mol/L组作用最强;高浓度(10-5mol/L)则抑制EPCs的上述功能。辛伐他汀促增殖作用随时间延长而增加,而粘附与体外血管形成能力则在48h作用最强。结论:辛伐他汀在较低浓度时显著增加了晚期EPCs的数量并改善其功能,高浓度则起抑制作用。Objective:To investigate the effect of simvastatin on the quantity and function of late endothelial progenitor cells(EPCs)from human umbilical cord blood in vitro.Method:Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation and cultured on fibronectin-coated culture flask, then the cells cultured in EGM-2MV medium differentiated into late EPCs which were identified by flow cytometry and fluorescent staining.The good growth state of late EPCs of second generation were treated with simvastatin in a series of concentrations(10-8,10-7,10-6,10-5 mol/L)for 24,48,72hand the proliferation,adhesion and tubule-formation activity of EPCs were analyzed by CCK-8reagent kit,adhesive assay and tubule-formation assay respectively.Result:Late EPCs were isolated and cultured successfully from human umbilical cord blood.Com- pared with control group,lower concentrations of simvastatin(10-8,10-7,10-6 mol/L)significantly promoted cell proliferation,adhesion and in vitro vascularization and the concentration of 10-7 mol/L was the most prominent,while higher concentration(1 0-5 mol/L)inhibited the functions above.Simvastatin increased the proliferative capacity of EPCs in a time dependent manner,but adhesion and tubule-formation activity were the most prominent at 48h.Conclusion:Simvastatin enhances the count of late EPCs and improve cell functions remarkably at lower concentrations,while higher concentration elicit inhibitory effects.
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